Incidental findings and ancillary-care obligations.
Recent work on incidental findings, concentrating on the difficult problems posed by the ambiguous results often generated by high-tech medicine, has proceeded largely independently from recent work on medical researchers' ancillary-care obligations, the obligations that researchers have to deal with diseases or conditions besides the one(s) under study. This paper contends that the two topics are morally linked, and specifically that a sound understanding of ancillary-care obligations will center them on incidental findings. The paper sets out and defends an understanding of ancillary-care obligations, which is based on the idea that when participants signed up for a study they may -- independently of their beliefs and expectations and of those of the researchers -- be taken to have partially entrusted certain aspects of their health into the researchers' hands. This partial entrustment model of ancillary-care obligations, in turn, has substantive implications for how to deal ethically with incidental findings; for instance, it suggests that researchers have no moral obligation to hunt for incidental findings.


Parental age and risk of autism spectrum disorders in a Finnish national birth cohort.
Aim of the study was to examine the associations between parental age and autism spectrum disorders (ASD). Data were based on the FIPS-A (Finnish Prenatal Study of Autism and Autism Spectrum Disorders), a case-control study with a total of 4,713 cases with childhood autism (n = 1,132), Asperger's syndrome (n = 1,785) or other pervasive developmental disorder (PDD) (n = 1,796), which were ascertained from the Finnish Hospital Discharge Register. Controls were selected from the Finnish Medical Birth Register. Conditional logistic regression models were used for statistical analyses. Advanced paternal age (35-49 years) was associated with childhood autism in offspring, whereas advanced maternal age was associated with both Asperger's syndrome and PDD in offspring (35 years or more and 40 years or more, respectively). Teenage motherhood (19 years or less) was associated with PDD in offspring. The main finding was that maternal and paternal ages were differentially associated with ASD subtypes. In addition to advanced parental age, teenage pregnancy seems to incur a risk for PDD in offspring.


Preoperative parathyroid localization by superimposed iodine-131 toluidine blue and technetium-99m pertechnetate imaging.
A new parathyroid scintigraphic localization study by a dual radioisotope technique using radioiodinated toluidine blue (RTB) for the parathyroids and 99mTc for thyroid imaging is presented. A simple RTB labeling procedure achieving 99% tagging of the 131I-TB was used. The RTB was found to be a highly specific parathyroid radiotracer, consequently enabling superimposition of the delineated thyroid gland over the RTB avid parathyroid foci without a need for subtraction of the thyroid or vascular background. Forty-six patients with primary hyperparathyroidism underwent scintigraphic study prior to cervical (41 patients) or mediastinal (5 patients) exploration and 67 pathological parathyroid glands (34 adenomas and 33 hyperplasias) were excised. On follow-up, serum calcium level returned to normal in all patients. Correlation of the scintigraphic results with the surgical findings disclosed a sensitivity of 93%, with a specificity of 80% and an overall accuracy of 87%. This new simplified and specific RTB scintigraphic method justifies its use as a routine procedure for preoperative parathyroid scintigraphic localization in primary hyperparathyroidism.


Mechanisms of oncogenic chromosomal translocations.
Chromosome translocations are caused by inappropriate religation of two DNA double-strand breaks (DSBs) in heterologous chromosomes. These DSBs can be generated by endogenous or exogenous sources. Endogenous sources of DSBs leading to translocations include inappropriate recombination activating gene (RAG) or activation-induced deaminase (AID) activity during immune receptor maturation. Endogenous DSBs can also occur at noncanonical DNA structures or at collapsed replication forks. Exogenous sources of DSBs leading to translocations include ionizing radiation (IR) and cancer chemotherapy. Spatial proximity of the heterologous chromosomes is also important for translocations. While three distinct pathways for DNA DSB repair exist, mounting evidence supports alternative nonhomologous end joining (aNHEJ) as the predominant pathway through which the majority of translocations occur. Initiated by poly (ADP-ribose) polymerase 1 (PARP1), aNHEJ is utilized less frequently in DNA DSB repair than other forms of DSB repair. We recently found that PARP1 is essential for chromosomal translocations to occur and that small molecule PARP1 inhibitors, already in clinical use, can inhibit translocations generated by IR or topoisomerase II inhibition. These data confirm the central role of PARP1 in aNHEJ-mediated chromosomal translocations and raise the possibility of using clinically available PARP1 inhibitors in patients who are at high risk for secondary oncogenic chromosomal translocations.


Differences in performance between Oticon MultiFocus Compact and ReSound BT2-E hearing aids.
Differences in performance were evaluated between binaural fittings of the Oticon MultiFocus (MF) and ReSound BT2-E on 25 hearing-impaired subjects across two sites. Subjects were initially fit using each manufacturer's algorithm and adjustments were made at 1 week based on subjects' responses to diary questions. Performance was assessed after a 4- to 6-week trial period with each hearing aid set using the Speech Perception in Noise (SPIN) test administered at 50, 65, and 80 dB SPL, the Abbreviated Profile of Hearing Aid Benefit (APHAB) questionnaire, loudness judgments of female connected discourse at 65 and 80 dB SPL, and an overall preference selection. The MF yielded significantly better SPIN scores at 50 and 65 dB SPL, while the BT2-E yielded a significantly better score at 80 dB SPL. No statistically significant differences were found in the APHAB benefit scores between the hearing aid sets, but both sets were significantly better than the subjects' own hearing aids on three of the four subscales. The MF produced slightly higher mean loudness judgments at both input levels than the BT2-E. Finally, 12 subjects preferred the BT2-E, 10 subjects preferred the MF, and three subjects stated no preference. The results are discussed in terms of audiogram effects on preference and effects of differences in signal processing approaches between the devices.


In Silico Discovery of Candidate Drugs against Covid-19.
Previous studies reported that Angiotensin converting enzyme 2 (ACE2) is the main cell receptor of SARS-CoV and SARS-CoV-2. It plays a key role in the access of the virus into the cell to produce the final infection. In the present study we investigated in silico the basic mechanism of ACE2 in the lung and provided evidences for new potentially effective drugs for Covid-19. Specifically, we used the gene expression profiles from public datasets including The Cancer Genome Atlas, Gene Expression Omnibus and Genotype-Tissue Expression, Gene Ontology and pathway enrichment analysis to investigate the main functions of ACE2-correlated genes. We constructed a protein-protein interaction network containing the genes co-expressed with ACE2. Finally, we focused on the genes in the network that are already associated with known drugs and evaluated their role for a potential treatment of Covid-19. Our results demonstrate that the genes correlated with ACE2 are mainly enriched in the sterol biosynthetic process, Aryldialkylphosphatase activity, adenosylhomocysteinase activity, trialkylsulfonium hydrolase activity, acetate-CoA and CoA ligase activity. We identified a network of 193 genes, 222 interactions and 36 potential drugs that could have a crucial role. Among possible interesting drugs for Covid-19 treatment, we found Nimesulide, Fluticasone Propionate, Thiabendazole, Photofrin, Didanosine and Flutamide.


Innovative Imaging Techniques Used to Evaluate Borderline-Resectable Pancreatic Adenocarcinoma.
A diagnosis of pancreatic cancer carries a 5-y survival rate of less than 10%. Furthermore, the detection of pancreatic cancer occurs most often in later stages of the disease due to its location in the retroperitoneum and lack of symptoms (in most cases) until tumors become more advanced. Once diagnosed, cross-sectional imaging techniques are heavily utilized to determine the tumor stage and the potential for surgical resection. However, a major determinant of resectability is the extent of local vascular involvement of the mesenteric vessels and critical tributaries; current imaging techniques have limited capacity to accurately determine vascular involvement. Surrounding inflammation and fibrosis can be difficult to discriminate from viable tumor, making determination of the degree of vascular involvement unreliable. New innovations in fluorescence and optoacoustic imaging techniques may overcome these limitations and make determination of resectability more accurate. These imaging modalities are able to more clearly discern between viable tumor tissue and non-neoplastic inflammation or desmoplasia, allowing clinicians to more reliably characterize vascular involvement and develop individualized treatment plans for patients. This review will discuss the current imaging techniques used to diagnose pancreatic cancer, the barriers that current techniques raise to accurate staging, and novel fluorescence and optoacoustic imaging techniques that may provide more accurate clinical staging of pancreatic cancer.


Identical genomic footprints of the adenovirus EIIa promoter are detected before and after EIa induction.
Genomic DNase I footprinting was used to compare specific protein binding to the adenovirus type 5 early, EIa-inducible, EIIa promoter. Identical protection patterns of the promoter region were observed whether EIIa transcription was undetectable or fully induced. These results suggest that EIa-mediated transcriptional induction does not increase binding of limiting transcription factors to the promoter but rather that transactivation results from the proper interactions between factors already bound to their cognate sequences.


Impaired neostigmine antagonism of pancuronium during enflurane anaesthesia in man.
We have compared the rates of recovery of pancuronium-induced neuromuscular blockade after administration of neostigmine during anaesthesia maintained with nitrous oxide and intermittent narcotics, halothane or enflurane. Thirty patients were studied in whom anaesthesia was maintained with 70% nitrous oxide in oxygen with: fentanyl or thiopentone, halothane (0.55-0.65% end-tidal), or enflurane (1.3-1.4% end-tidal). Muscle twitch response was measured using train-of-four stimulation. Pancuronium 3 mg/70 kg was antagonized with neostigmine 2.5 mg/70 kg at 10% spontaneous recovery of the first twitch of the train compared with control. There were no significant differences between the times to 10% spontaneous recovery of the first twitch or the rates of train-of-four recovery after neostigmine when the narcotic and halothane groups were compared. However, enflurane anaesthesia, in comparison with fentanyl, was associated with a significant increase in the time to 10% recovery (57.9 +/- 6.0 min v. 35.4 +/- 7.0 min) and with decreases in the train-of-four recoveries at 4, 5, 10, 20 and 30 min after neostigmine. We conclude that, under the conditions of this study, the antagonism of pancuronium-induced neuromuscular blockade with neostigmine was impaired during enflurane but not halothane anaesthesia.


[Neuropsychiatric aspects of diagnostics of nonepileptic seizures].
The outcome of nonepileptic seizures (NES) is usually poor. The early recognition suggests better prognosis. This study is undertaken to examine the value of neuropsychiatric method for making accurate diagnosis of the psychiatric disorder that underlies psychogenic NES. Multidisciplinary approach to clinical assessment of such patients with chronic or resistant course of seizure disorder was used. Out of 6 patients, who went through the neuropsychiatric evaluation, two were admitted with a diagnosis of status epilepticus, three--with acute episodes of repetitive seizures and one--with diagnoses of bettolepsy. The results demonstrated that in four cases seizures were the manifestation of dissociative disorder (F 44), in one--of somatic form autonomic dysfunction (F 45.3) and the last patient had delirium due to epilepsy. All of the four patients with the past history of epilepsy were treated with antiepileptic drugs (AED). In two cases diagnosis of epilepsy and AED therapy were withdrawn. In the remaining cases we supposed the coexistence of epilepsy and NES. Our data suggests that incorporating of neuropsychiatric methods into the diagnostic battery for people with suspected and/or intractable epilepsy help in distinguishing NES and epilepsy as well as in revealing the underlying psychic disorder, which leads to formulation specific treatment program.


Longitudinal study of AMH variations in 122 Adolescents and Young Adults (AYA) and non-AYA lymphoma patients to evaluate the chemo-induced ovarian toxicity to further personalise fertility preservation counselling.
What is the influence of age and chemotherapy regimen on the longitudinal blood anti-Müllerian hormone (AMH) variations in a large series of adolescents and young adult (AYA) (15-24 years old) and non-AYA (25-35 years old) lymphoma patients? In case of alkylating regimen treatment, there was a deep and sustained follicular depletion in AYA as well as non-AYA patients; however in both groups, the ovarian toxicity was extremely low in cases of non-alkylating treatments. AMH is now well-recognised to be a real-time indicator of ovarian follicular depletion and recovery in women treated by chemotherapy. Its longitudinal variations may discriminate between highly and minimally toxic protocols regarding ovarian function. It has been shown, in different cancer types, that age, type of chemotherapy regimen and pre-treatment AMH levels are the main predictors of ovarian recovery. Large studies on longitudinal AMH variations under chemotherapy in lymphoma patients are few but can provide the opportunity to assess the degree of follicle loss at a young age. This prospective cohort study was conducted in the Fertility Observatory of the Lille University Hospital. Data were collected between 2007 and 2016. Non-Hodgkin or Hodgkin lymphoma patients (n = 122) between 15 and 35 years old were prospectively recruited before commencing chemotherapy. Patients were treated either by a non-alkylating protocol (ABVD group; n = 67) or by an alkylating regimen (alkylating group; n = 55). Serial AMH measurements were performed at baseline (AMH0), 15 days after the start of chemotherapy (AMH1), 15 days before the last chemotherapy cycle (AMH2), and at time 3, 6, 9, 12, 18 and 24 months from the end of chemotherapy. The whole study population was divided into two groups according to age: AYA (15-24; n = 65) and non-AYA (25-35; n = 57). All patients received a once monthly GnRH agonist injection during the whole treatment period. A linear mixed model was used to account for the repeated measures of single patients. At baseline, non-AYA patients had higher BMI and lower AMH levels than AYA patients. All AYA and non-AYA patients having received ABVD protocols had regular cycles at 12 months of follow-up. In case of alkylating regimens, amenorrhoea was more frequent in non-AYA patients than in AYA patients at 12 months (37% vs 4%, P = 0.011) and at 24 months (24% vs 4%, P = 0.045). We distinguished a similar depletion phase from AMH0 to AMH2 between ABVD and alkylating groups but significantly different recovery phases from AMH2 to AMH + 24 months. AMH recovery was fast and complete in case of ABVD protocols whatever the age: AMH reached pre-treatment values as soon as the 6th month of follow-up in the AYA group (mean (95% CI) in log AMH M0 vs M6: 3.07 (2.86 to 3.27) vs 3.05 (2.78 to 3.31), P = 1.00) and in the non-AYA group (mean (95% CI) in log AMH M0 vs M6: 2.73 (2.40 to 3.05) vs 2.47 (2.21 to 2.74), P = 1.00). In contrast, no patients from the alkylating group returned to pre-treatment AMH values whatever the age of patients (AYA or non-AYA). Moreover, none of the AMH values post-chemotherapy in the non-AYA group were significantly different from AMH2. Conversely in the AYA group, AMH levels from 6 months (mean (95% CI) in log AMH: 1.79 (1.47 to 2.11), P < 0.001) to 24 months (mean (95% CI) in log AMH: 2.16 (1.80 to 2.52), P ≤ 0.001) were significantly higher than AMH2 (mean (95% CI) in log AMH: 1.13 (0.89 to 1.38)). Considering the whole study population (AYA and non-AYA), pre-treatment AMH levels influenced the pattern of the AMH variation both in alkylating and ABVD protocols (interaction P-value = 0.005 and 0.043, respectively). Likewise, age was significantly associated with the pattern of the recovery phase but only in the alkylating group (interaction P-value =0.001). BMI had no influence on the AMH recovery phase whatever the protocol (interaction P-value = 0.98 in alkylating group, 0.72 in ABVD group). There was a large disparity in subtypes of protocols in the alkylating group. The average duration of chemotherapy for patients treated with alkylating protocols was longer than that for patients treated with ABVD. These results make it possible to develop strategies for fertility preservation according to age and type of protocol in a large series of young lymphoma patients. In addition, it was confirmed that young age does not protect against ovarian damage caused by alkylating agents. This work was supported by Agence Régionale de Santé Hauts de France and Agence Onco Hauts-de-France who provided finances for AMH dosages (n° DOS/SDES/AR/FIR/2019/282). There are no competing interests. DC-2008-642 and CNIL DEC2015-112.


Differential sensitivity to detergents of actin cytoskeleton from nerve endings.
Detergent-resistant membranes (DRM), an experimental model used to study lipid rafts, are typically extracted from cells by means of detergent treatment and subsequent ultracentrifugation in density gradients, Triton X-100 being the detergent of choice in most of the works. Since lipid rafts are membrane microdomains rich in cholesterol, depletion of this component causes solubilization of DRM with detergent. In previous works from our group, the lack of effect of cholesterol depletion on DRM solubilization with Triton X-100 was detected in isolated rat brain synaptosomes. In consequence, the aim of the present work is to explore reasons for this observation, analyzing the possible role of the actin cytoskeleton, as well as the use of an alternative detergent, Brij 98, to overcome the insensitivity to Triton X-100 of cholesterol-depleted DRM. Brij 98 yields Brij-DRM that are highly dependent on cholesterol, since marker proteins (Flotillin-1 and Thy-1), as well as actin, appear solubilized after MCD treatment. Pretreatment with Latrunculin A results in a significant increase in Flotillin-1, Thy-1 and actin solubilization by Triton X-100 after cholesterol depletion. Studies with transmission electron microscopy show that combined treatment with MCD and Latrunculin A leads to a significant increase in solubilization of DRM with Triton X-100. Thus, Triton-DRM resistance to cholesterol depletion can be explained, at least partially, thanks to the scaffolding action of the actin cytoskeleton, without discarding differential effects of Brij 98 and Triton X-100 on specific membrane components. In conclusion, the detergent of choice is important when events that depend on the actin cytoskeleton are going to be studied.


The Efficacy and Safety of Gabapentinoids in Total Joint Arthroplasty: Systematic Review and Direct Meta-Analysis.
Gabapentinoids are commonly used as an adjunct to traditional pain management strategies after total joint arthroplasty (TJA). The purpose of this study is to evaluate the efficacy and safety of gabapentinoids in primary TJA to support the combined clinical practice guidelines of the American Association of Hip and Knee Surgeons, American Academy of Orthopaedic Surgeons, Hip Society, Knee Society, and the American Society of Regional Anesthesia and Pain Management. The MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials were searched for studies published prior to November 2018 on gabapentinoids in TJA. All included studies underwent qualitative and quantitative homogeneity testing followed by a systematic review and direct comparison meta-analysis to assess the efficacy and safety of gabapentinoids. In total, 384 publications were critically appraised to provide 13 high-quality studies regarded as the best available evidence for analysis. In the perioperative period prior to discharge, pregabalin reduces postoperative opioid consumption, but gabapentinoids do not reduce postoperative pain. After discharge, gabapentin does not reduce postoperative pain or opioid consumption, but pregabalin reduces both postoperative pain and opioid consumption. Moderate evidence supports the use of pregabalin in TJA to reduce postoperative pain and opioid consumption. Gabapentinoids should be used with caution, however, as they may lead to an increased risk of sedation and respiratory depression especially when combined with other central nervous system depressants such as opioids.


The effect of the dispersal kernel on isolation-by-distance in a continuous population.
Under models of isolation-by-distance, population structure is determined by the probability of identity-by-descent between pairs of genes according to the geographic distance between them. Well established analytical results indicate that the relationship between geographical and genetic distance depends mostly on the neighborhood size of the population which represents a standardized measure of gene flow. To test this prediction, we model local dispersal of haploid individuals on a two-dimensional landscape using seven dispersal kernels: Rayleigh, exponential, half-normal, triangular, gamma, Lomax and Pareto. When neighborhood size is held constant, the distributions produce similar patterns of isolation-by-distance, confirming predictions. Considering this, we propose that the triangular distribution is the appropriate null distribution for isolation-by-distance studies. Under the triangular distribution, dispersal is uniform over the neighborhood area which suggests that the common description of neighborhood size as a measure of an effective, local panmictic population is valid for popular families of dispersal distributions. We further show how to draw random variables from the triangular distribution efficiently and argue that it should be utilized in other studies in which computational efficiency is important.


[The molecular scenarios of the consolidation of long-term memory].
Long-term memory consolidation is a critical event in the transition of short-lasting experiences into durable modifications of behaviour. Present article focuses on the problem of molecular bases of this process. It starts with a brief review of biochemical and pharmacological data demonstrating a universal dependence of long-term memory on gene expression in the brain. Some of the experimental studies of immediate early gene expression in the brain during learning are described in the second part of the article. A hypothesis is discussed according to which consolidation of long-term memory employ the same biphasic molecular cascade of gene expression that is used for cell growth and differentiation during development.


Role of histamine H1 and H2 receptor antagonists in the prevention of intimal thickening.
Vascular smooth muscle cell migration to the intima from the media and proliferation in the intima play key roles in atherosclerosis and restenosis after coronary angioplasty. Histamine released from adherent platelets at the injured artery and from mast cells in atheromas has stimulant actions on both smooth muscle cell migration and proliferation, and histamine receptor antagonists abolish the effect of histamine in vitro. The aim of this study was to examine the effect of histamine receptor antagonists on intimal thickening. Endothelial injury in the mouse femoral artery was induced by a photochemical reaction between localized irradiation by green light and intravenously administered rose bengal. The histamine H1 receptor antagonist, diphenhydramine, at a dose of 30 mg/kg or the histamine H2 receptor antagonist, cimetidine, at a dose of 200 mg/kg was intraperitoneally administered to mice for 21 days after endothelial injury. Twenty-one days after endothelial injury, morphometric analysis was performed to measure the cross-sectional areas of the intima and media. Diphenhydramine significantly reduced the intimal area to 1.1 +/- 0.3 (x 10(-3) mm2) compared with the value in the control group, which was 6.2 +/- 1.4 (x 10(-3) mm2), but cimetidine (5.5 +/- 1.9, x 10(-3) mm2) did not. Similarly, the ratio of intimal area to medial area in the diphenhydramine-treated group but not in the cimetidine-treated group was significantly reduced (83%). In the in vitro study, cimetidine inhibited neither proliferation nor migration of mouse vascular smooth muscle cells stimulated by platelet-derived growth factor (PDGF). In contrast, diphenhydramine significantly (P < 0.05) inhibited proliferation in a dose-dependent manner, but did not inhibit migration. These results suggest that diphenhydramine, a histamine H1 receptor antagonist, reduced the formation of intimal hyperplasia, at least in part due to inhibition of cell proliferation. However, cimetidine, a histamine H2 receptor antagonist, was ineffective. Histamine may play a key role in intimal thickening, in part via histamine H1 receptors in this model.


Effects of cadmium on the energy metabolism of isolated hepatocytes: its relationship with the nonviability of isolated hepatocytes caused by cadmium.
The effects of cadmium on the energy metabolism of mitochondria were studied with isolated hepatocytes and rat liver mitochondria. It was found that cadmium inhibited the respiration of both isolated hepatocytes and mitochondria and decreased the ATP content of isolated hepatocytes. This inhibition of energy metabolism of mitochondria was highly related to the nonviability of isolated hepatocytes caused by cadmium. The site of electron transport of the mitochondrial respiratory chain blocked by cadmium was located between cytochrome b and flavo-proteins. The uncoupling effects of mitochondrial oxidative phosphorylation caused by cadmium may have resulted from changes in the fluidity and permeability of the mitochondrial membrane.


Mitochondrial oxidant stress increases PDE5 activity in persistent pulmonary hypertension of the newborn.
In the pulmonary vasculature, phosphodiesterase-5 (PDE5) degrades cGMP and inhibits nitric oxide-mediated, cGMP-dependent vasorelaxation. We previously reported that ventilation with 100% O2 increased PDE5 activity in pulmonary arteries (PAs) of pulmonary hypertension lambs (PPHN) more than in control lambs. In the present study, PA smooth muscle cells (PASMCs) from PPHN lambs had increased basal PDE5 activity, decreased cGMP-responsiveness to NO, and increased mitochondrial matrix oxidant stress compared to control PASMC. Hyperoxia (24 h) increased PDE5 activity and mitochondrial matrix oxidant stress above baseline to a similar degree in PPHN and control PASMC. Mitochondrially targeted catalase decreased PDE5 activity at baseline and after hyperoxia in PPHN PASMC. Similarly, catalase treatment of PPHN lambs ventilated with 100% O2 decreased PDE5 activity and increased cGMP in PA. We conclude that baseline PDE5 activity and oxidative stress is increased in PPHN PASMC, and scavenging H2O2 is sufficient to block oxidant-mediated increases in PDE5 activity in PPHN.


Choledochopancreatoduodenal fistula caused by duodenal ulceration. A case report.
A penetrating duodenal ulcer may occasionally erode into the common bile duct and form a choledochoduodenal fistula. Such a fistula occurring simultaneously with a pancreatic duodenal fistula is reported. The presenting features of these fistulas are those of the ulcer and confirmation of the fistula may be difficult, although use of endoscopic retrograde cholangiopancreatography has greatly facilitated their diagnosis. In this case both fistulas could be cannulated through the base of the ulcer. The majority of these fistulas heal spontaneously with intensive medical management. The remainder require surgery, and a conservative approach avoiding direct interference with the fistula should be adopted. Drainage procedures are rarely required and once closed the fistulas usually cause no further problem.


Cannabimimetic fatty acid derivatives in cancer and inflammation.
Evidence for the role of the cannabimimetic fatty acid derivatives (CFADs), i.e. anandamide (arachidonoylethanolamide, AEA), 2-arachidonoylglycerol (2-AG) and palmitoylethanolamide (PEA), in the control of inflammation and of the proliferation of tumor cells is reviewed here. The biosynthesis of AEA, PEA, or 2-AG can be induced by stimulation with either Ca(2+) ionophores, lipopolysaccharide, or platelet activating factor in macrophages, and by ionomycin or antigen challenge in rat basophilic leukemia (RBL-2H3) cells (a widely used model for mast cells). These cells also inactivate CFADs through re-uptake and/or hydrolysis and/or esterification processes. AEA and PEA modulate cytokine and/or arachidonate release from macrophages in vitro, regulate serotonin secretion from RBL-2H3 cells, and are analgesic in some animal models of inflammatory pain. However, the involvement of endogenous CFADs and cannabinoid CB(1) and CB(2) receptors in these effects is still controversial. In human breast and prostate cancer cells, AEA and 2-AG, but not PEA, potently inhibit prolactin and/or nerve growth factor (NGF)-induced cell proliferation. Vanillyl-derivatives of anandamide, such as olvanil and arvanil, exhibit even higher anti-proliferative activity. These effects are due to suppression of the levels of the 100 kDa prolactin receptor or of the high affinity NGF receptors (trk), are mediated by CB(1)-like cannabinoid receptors, and are enhanced by other CFADs. Inhibition of adenylyl cyclase and activation of mitogen-activated protein kinase underlie the anti-mitogenic actions of AEA. The possibility that CFADs act as local inhibitors of the proliferation of human breast cancer is discussed here.


[The GABA-ergic system and brain edema].
It has been shown in rats with experimental toxic and traumatic edemas that picrotoxin (1 mg/kg) removes the antiedematous action of diazepam, phenazepam, phenibut and amizyl and reduces the action of phentolamine. When the dose of picrotoxin is minimized to 0.5 mg/kg such an effect is not observed. Prolonged daily administration of picrotoxin in a dose of 1 mg/kg results in the development of brain edema. It is recommended that GABA-positive drugs be included into a complex of treatment measures for edema.


Zinc status and serum testosterone levels of healthy adults.
Zinc deficiency is prevalent throughout the world, including the USA. Severe and moderate deficiency of zinc is associated with hypogonadism in men. However, the effect of marginal zinc deficiency on serum testosterone concentration is not known. We studied the relationship between cellular zinc concentrations and serum testosterone cross-sectionally in 40 normal men, 20 to 80 y of age. In four normal young men (27.5 +/- 0.5 y), we measured serum testosterone before and during marginal zinc deficiency induced by restricting dietary zinc intake. We also measured serum testosterone in nine elderly men (64 +/- 9 y) who were marginally zinc deficient before and after 3 to 6 mo of supplementation with 459 mumol/ d oral zinc administered as zinc gluconate. Serum testosterone concentrations were significantly correlated with cellular zinc concentrations in the cross-sectional study (lymphocyte zinc versus serum testosterone, r = 0.43, p = 0.006; granulocyte zinc versus serum testosterone, r = 0.30, p = 0.03). Dietary zinc restriction in normal young men was associated with a significant decrease in serum testosterone concentrations after 20 weeks of zinc restriction (baseline versus post-zinc restriction mean +/- SD, 39.9 +/- 7.1 versus 10.6 +/- 3.6 nmol/L, respectively; p = 0.005). Zinc supplementation of marginally zinc-deficient normal elderly men for six months resulted in an increase in serum testosterone from 8.3 +/- 6.3 to 16.0 +/- 4.4 nmol/L (p = 0.02). We conclude that zinc may play an important role in modulating serum testosterone levels in normal men.


The diagnostic accuracy of biomarkers for diagnosis of primary biliary cholangitis (PBC) in anti-mitochondrial antibody (AMA)-negative PBC patients: a review of literature.
Primary biliary cholangitis (PBC), also known as primary biliary cirrhosis, is an autoimmune disease of the liver characterized by anti-mitochondrial antibodies (AMA) in 90%-95% of patients. The aim of this study was to evaluate the diagnostic value of several serum biomarkers in patients with PBC but negative for AMA. Some antinuclear antibodies (ANA) pattern, detected by indirect immunofluorescence (IIF), such as multiple nuclear dot (MND) and rim-like patterns are well-known to be specific for PBC. The corresponding nuclear antigens are the components of the nuclear pore complex (Gp210 for rim-like pattern) and Sp100, PML proteins (for MND pattern) detectable by immunoblotting and ELISA methods. More recently, new biomarkers have been evaluated in order to improve the diagnostic sensitivity, such as kelch-like 12 (KLHL12) and hexokinase-1. Considering these different serum biomarkers, studies evaluating their diagnostic role in AMA-negative PBC patients compared to AMA-positive ones and controls were included in this review. Pooled sensitivity and specificity were 37% and 85%, respectively. The overall PPV and NPV mean values were 45% and 83%. Even if all biomarkers are very specific for PBC, the overall sensitivity was poor and therefore is necessary to identify a marker with a greater sensitivity for PBC in AMA-negative patients.


Measures of patient radiation exposure during endoscopic retrograde cholangiography: beyond fluoroscopy time.
To determine whether fluoroscope time is a good predictor of patient radiation exposure during endoscopic retrograde cholangiopancreatography. This is a prospective observational study of consecutive patients undergoing endoscopic retrograde cholangiopancreatography in a tertiary care setting. Data related to radiation exposure were collected. The following measures were obtained: Fluoroscopy time (FT), dose area product (DAP) and dose at reference point (DOSERP). Coefficients of determination were calculated to analyze the correlation between FT, DAP and DOSRP. Agreement between FT and DAP/DOSRP was assessed using Bland Altman plots. Four hundred sixty-three data sets were obtained. Fluoroscopy time average was 7.3 min. Fluoroscopy related radiation accounted for 86% of the total DAP while acquisition films related radiation accounted for 14% of the DAP. For any given FT there are wide ranges of DAP and DOSERP and the variability in both increases as fluoroscopy time increases. The coefficient of determination (R(2)) on the non transformed data for DAP and DOSERP versus FT were respectively 0.416 and 0.554. While fluoroscopy use was the largest contributor to patient radiation exposure during endoscopic retrograde cholangiography (ERCP), there is a wide variability in DAP and DOSERP that is not accounted for by FT. DAP and DOSERP increase in variability as FT increases. This translates into poor accuracy of FT in predicting DAP and DOSERP at higher radiation doses. DAP and DOSERP in addition to FT should be adopted as new ERCP quality measures to estimate patient radiation exposure.


painACTION-back pain: a self-management website for people with chronic back pain.
To determine whether an interactive self-management Website for people with chronic back pain would significantly improve emotional management, coping, self-efficacy to manage pain, pain levels, and physical functioning compared with standard text-based materials. The study utilized a pretest-posttest randomized controlled design comparing Website (painACTION-Back Pain) and control (text-based material) conditions at baseline and at 1-, 3, and 6-month follow-ups. Two hundred and nine people with chronic back pain were recruited through dissemination of study information online and at a pain treatment clinic. The 6-month follow-up rates for the Website and control groups were 73% and 84%, respectively. Measures were based on the recommendations of the Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials and included measures of pain intensity, physical functioning, emotional functioning, coping, self-efficacy, fear-avoidance, perceived improvement with treatment, self-efficacy, and catastrophizing. Compared with controls, painACTION-Back Pain participants reported significantly: 1) lower stress; 2) increased coping self-statements; and 3) greater use of social support. Comparisons between groups suggested clinically significant differences in current pain intensity, depression, anxiety, stress, and global ratings of improvement. Among participants recruited online, those using the Website reported significantly: 1) lower "worst" pain; 2) lower "average" pain; and 3) increased coping self-statements, compared with controls. Participants recruited through the pain clinic evidenced no such differences. An online self-management program for people with chronic back pain can lead to improvements in stress, coping, and social support, and produce clinically significant differences in pain, depression, anxiety, and global rates of improvement.


The impact of restaurant consumption among US adults: effects on energy and nutrient intakes.
To examine the effect of fast-food and full-service restaurant consumption on adults' energy intake and dietary indicators. Individual-level fixed-effects regression model estimation based on two different days of dietary intake data was used. Parallel to the rising obesity epidemic in the USA, there has been a marked upward trend in total energy intake derived from food away from home. The full sample included 12 528 respondents aged 20-64 years who completed 24 h dietary recall interviews for both day 1 and day 2 in the National Health and Nutrition Examination Survey (NHANES) 2003-2004, 2005-2006, 2007-2008 and 2009-2010. Fast-food and full-service restaurant consumption, respectively, was associated with an increase in daily total energy intake of 813·75 kJ (194·49 kcal) and 858·04 kJ (205·21 kcal) and with higher intakes of saturated fat (3·48 g and 2·52 g) and Na (296·38 mg and 451·06 mg). Individual characteristics moderated the impacts of restaurant food consumption with adverse impacts on net energy intake being larger for black adults compared with their white and Hispanic counterparts and greater for middle-income v. high-income adults. Adults' fast-food and full-service restaurant consumption was associated with higher daily total energy intake and poorer dietary indicators.


Reversible patterning and actuation of hydrogels by electrically assisted ionoprinting.
The ability to pattern, structure, re-shape and actuate hydrogels is important for biomimetics, soft robotics, cell scaffolding and biomaterials. Here we introduce an 'ionoprinting' technique with the capability to topographically structure and actuate hydrated gels in two and three dimensions by locally patterning ions via their directed injection and complexation, assisted by electric fields. The ionic binding changes the local mechanical properties of the gel to induce relief patterns and, in some cases, evokes localized stress large enough to cause rapid folding. These ionoprinted patterns are stable for months, yet the ionoprinting process is fully reversible by immersing the gel in a chelator. The mechanically patterned hydrogels exhibit programmable temporal and spatial shape transitions, and serve as a basis for a new class of soft actuators that can gently manipulate objects both in air and in liquid solutions.


Cytotoxic effects of metal protoporphyrins in glioblastoma cells: roles of albumin, reactive oxygen species, and heme oxygenase-1.
We investigate the cytotoxic effect of metal protoporphyrins including ferric protoporphyrin (FePP; hemin), cobalt protoporphyrin (CoPP), and tin protoporphyrin (SnPP) in glioblastoma cells C6 and GBM8401. Data of MTT assay show that FePP and CoPP, but not SnPP, significantly reduce the viability of glioma cells C6 and GBM8401 in the absence of serum. In the condition with fetal bovine serum (FBS) or bovine serum albumin (BSA), the cytotoxic effect of FePP and CoPP was completely inhibited. Binding of FePP, CoPP, and SnPP with BSA was examined via spectrophotometer analysis, and the protective effect of serum against FePP and CoPP-induced cell death was abolished by BSA depletion. A loss in the integrity of DNA with an occurrence of apoptotic events including DNA ladders, caspase 3 and PARP protein cleavage, and chromatin-condensed cells is observed in FePP-treated or CoPP-treated C6 cells. An increase in intracellular peroxide level was examined in FePP, but not CoPP, -treated C6 cells, and N-acetyl-l-cysteine (NAC) addition significantly protected C6 cells from FePP, but not CoPP, -induced cell death with reducing FePP-stimulated reactive oxygen species (ROS) production. Activation of extracellular regulated kinases (ERKs) and c-Jun-N-terminal kinases (JNKs) with an increase in the heme oxygenase-1 (HO-1) protein was observed in FePP-treated or CoPP-treated C6 cells in the absence of FBS or BSA, and adding JNKs inhibitor SP600125 (SP), but not ERKs inhibitor PD98059 (PD), significantly attenuated FePP-induced or CoPP-induced HO-1 protein expression in accordance with reducing JNKs protein phosphorylation. However, PD98059, SP600125, or transfection of C6 cells with antisense HO-1 oligonucleotides show no effect on the cytotoxicity elicited by FePP and CoPP in C6 cells. Effect of serum and BSA on the cytotoxicity of metal protoporphyrins in glioma cells is first demonstrated in the present study, and the roles of ROS, MAPKs, and HO-1 were elucidated.


Inventory Control System for a Healthcare Apparel Service Centre with Stockout Risk: A Case Analysis.
Based on the real-world inventory control problem of a capacitated healthcare apparel service centre in Hong Kong which provides tailor-made apparel-making services for the elderly and disabled people, this paper studies a partial backordered continuous review inventory control problem in which the product demand follows a Poisson process with a constant lead time. The system is controlled by an (Q,r) inventory policy which incorporate the stockout risk, storage capacity, and partial backlog. The healthcare apparel service centre, under the capacity constraint, aims to minimize the inventory cost and achieving a low stockout risk. To address this challenge, an optimization problem is constructed. A real case-based data analysis is conducted, and the result shows that the expected total cost on an order cycle is reduced substantially at around 20% with our proposed optimal inventory control policy. An extensive sensitivity analysis is conducted to generate additional insights.


Disruption of MAGI2-RapGEF2-Rap1 signaling contributes to podocyte dysfunction in congenital nephrotic syndrome caused by mutations in MAGI2.
The essential role of membrane associated guanylate kinase 2 (MAGI2) in podocytes is indicated by the phenotypes of severe glomerulosclerosis of both MAGI2 knockout mice and in patients with congenital nephrotic syndrome (CNS) caused by mutations in MAGI2. Here, we show that MAGI2 forms a complex with the Rap1 guanine nucleotide exchange factor, RapGEF2, and that this complex is lost when expressing MAGI2 CNS variants. Co-expression of RapGEF2 with wild-type MAGI2, but not MAGI2 CNS variants, enhanced activation of the small GTPase Rap1, a central signaling node in podocytes. In mice, podocyte-specific RapGEF2 deletion resulted in spontaneous glomerulosclerosis, with qualitative glomerular features comparable to MAGI2 knockout mice. Knockdown of RapGEF2 or MAGI2 in human podocytes caused similar reductions in levels of Rap1 activation and Rap1-mediated downstream signaling. Furthermore, human podocytes expressing MAGI2 CNS variants show severe abnormalities of cellular morphology and dramatic loss of actin cytoskeletal organization, features completely rescued by pharmacological activation of Rap1 via a non-MAGI2 dependent upstream pathway. Finally, immunostaining of kidney sections from patients with congenital nephrotic syndrome and MAGI2 mutations showed reduced podocyte Rap1-mediated signaling. Thus, MAGI2-RapGEF2-Rap1 signaling is essential for normal podocyte function. Hence, disruption of this pathway is an important cause of the renal phenotype induced by MAGI2 CNS mutations.


Association between the skin microbiome and MHC class II diversity in an amphibian.
Microbiomes play an important role in determining the ecology and behaviour of their hosts. However, questions remain pertaining to how host genetics shape microbiomes, and how microbiome composition influences host fitness. We explored the effects of geography, evolutionary history and host genetics on the skin microbiome diversity and structure in a widespread amphibian. More specifically, we examined the association between bacterial diversity and composition and the major histocompatibility complex class II exon 2 diversity in 12 moor frog (Rana arvalis) populations belonging to two geographical clusters that show signatures of past and ongoing differential selection. We found that while bacterial alpha diversity did not differ between the two clusters, MHC alleles/supertypes and genetic diversity varied considerably depending on geography and evolutionary history. Bacterial alpha diversity was positively correlated with expected MHC heterozygosity and negatively with MHC nucleotide diversity. Furthermore, bacterial community composition showed significant variation between the two geographical clusters and between specific MHC alleles/supertypes. Our findings emphasize the importance of historical demographic events on hologenomic variation and provide new insights into how immunogenetic host variability and microbial diversity may jointly influence host fitness with consequences for disease susceptibility and population persistence.


Cancer Risk in Subsolid Nodules in the National Lung Screening Trial.
Background Subsolid pulmonary nodules, comprising pure ground-glass nodules (GGNs) and part-solid nodules (PSNs), have a high risk of indolent malignancy. Lung Imaging Reporting and Data System (Lung-RADS) nodule management guidelines are based on expert opinion and lack independent validation. Purpose To evaluate Lung-RADS estimates of the malignancy rates of subsolid nodules, using nodules from the National Lung Screening Trial (NLST), and to compare Lung-RADS to the NELSON trial classification as well as the Brock University calculator. Materials and Methods Subsets of GGNs and PSNs were selected from the NLST for this retrospective study. A thoracic radiologist reviewed the baseline and follow-up CT images, confirmed that they were true subsolid nodules, and measured the nodules. The primary outcome for each nodule was the development of malignancy within the follow-up period (median, 6.5 years). Nodules were stratified according to Lung-RADS, NELSON trial criteria, and the Brock model. For analyses, nodule subsets were weighted on the basis of frequency in the NLST data set. Nodule stratification models were tested by using receiver operating characteristic curves. Results A total of 622 nodules were evaluated, of which 434 nodules were subsolid. At baseline, 304 nodules were classified as Lung-RADS category 2, with a malignancy rate of 3%, which is greater than the 1% in Lung-RADS (P = .004). The malignancy rate for GGNs smaller than 10 mm (two of 129, 1.3%) was smaller than that for GGNs measuring 10-19 mm (11 of 153, 6%) (P = .01). The malignancy rate for Lung-RADS category 3 was 14% (13 of 67), which is greater than the reported 2% in Lung-RADS (P < .001). The Brock model predicted malignancy better than Lung-RADS and the NELSON trial scheme (area under the receiver operating characteristic curve = 0.78, 0.70, and 0.67, respectively; P = .02 for Brock model vs NELSON trial scheme). Conclusion Subsolid nodules classified as Lung Imaging Reporting and Data System (Lung-RADS) categories 2 and 3 have a higher risk of malignancy than reported. The Brock risk calculator performed better than measurement-based classification schemes such as Lung-RADS. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Kauczor and von Stackelberg in this issue.


Functional specialization among members of Knickkopf family of proteins in insect cuticle organization.
Our recent study on the functional analysis of the Knickkopf protein from T. castaneum (TcKnk), indicated a novel role for this protein in protection of chitin from degradation by chitinases. Knk is also required for the laminar organization of chitin in the procuticle. During a bioinformatics search using this protein sequence as the query, we discovered the existence of a small family of three Knk-like genes (including the prototypical TcKnk) in the T. castaneum genome as well as in all insects with completed genome assemblies. The two additional Knk-like genes have been named TcKnk2 and TcKnk3. Further complexity arises as a result of alternative splicing and alternative polyadenylation of transcripts of TcKnk3, leading to the production of three transcripts (and by inference, three proteins) from this gene. These transcripts are named TcKnk3-Full Length (TcKnk3-FL), TcKnk3-5' and TcKnk3-3'. All three Knk-family genes appear to have essential and non-redundant functions. RNAi for TcKnk led to developmental arrest at every molt, while down-regulation of either TcKnk2 or one of the three TcKnk3 transcripts (TcKnk3-3') resulted in specific molting arrest only at the pharate adult stage. All three Knk genes appear to influence the total chitin content at the pharate adult stage, but to variable extents. While TcKnk contributes mostly to the stability and laminar organization of chitin in the elytral and body wall procuticles, proteins encoded by TcKnk2 and TcKnk3-3' transcripts appear to be required for the integrity of the body wall denticles and tracheal taenidia, but not the elytral and body wall procuticles. Thus, the three members of the Knk-family of proteins perform different essential functions in cuticle formation at different developmental stages and in different parts of the insect anatomy.


Subacute Oral Toxicity Evaluation of Freeze-Dried Powder of Locusta migratoria.
Novel food sources have enormous potential as nutritional supplements. For instance, edible insects are considered as an alternative food source due to their higher protein content; moreover, they are economically efficient reproducers and have high in nutritional value. In this study, we investigated the toxicity of the freeze-dried powder of Locusta migratoria (fdLM), known to contain rich proteins as well as fatty acids. The objective of the present study was to evaluate the subacute toxicity of fdLM in male and female Sprague-Dawley (SD) rats. The SD rats were divided into four groups based on the dosage of fdLM administered: dosage of 0 (vehicle control), 750, 1,500, and 3,000 mg/kg/day were administered for 28 days. Toxicological assessments including observations on food consumption, body and organ weights, clinical signs, mortality, ophthalmologic tests, urinalyses, hematologic tests, clinical chemistry tests, gross findings, and histopathology tests were performed. Clinical signs, urinalyses, hematology, serum biochemistry tests, and organ weight examinations revealed no fdLM-related toxicity. The no-observed-adverse-effect level for fdLM was higher than 3,000 mg/kg/day in rats of both sexes; therefore, fdLM, in conclusion, can be considered safe as an edible alternative human and animal food source material.


COX-2 inhibitor prevents the development of hyperalgesia induced by intrathecal NMDA or AMPA.
Cyclooxygenase (COX)-2 is constitutively expressed in the superficial dorsal horn of the spinal cord, but its role in the spinal cord is still unclear. We examined the effect of intrathecally administered NS398, a selective COX-2 inhibitor and indomethacin, an non-selective COX-1 and COX-2 inhibitor, on the development of thermal hyperalgesia induced by the activation of NMDA or AMPA receptors. Intrathecal injection of either NS398 or indomethacin equally blocked the development of thermal hyperalgesia induced by intrathecal injection of either NMDA or AMPA in a dose-dependent manner. These data suggest that COX-2 plays an important role in spinal thermal nociceptive transmission when neurons in the spinal cord are facilitated by NMDA or AMPA.


DNA binding properties of the Ah receptor in wild-type and variant mouse hepatoma cells.
Mouse hepatoma Hepa 1c1c7 cells and nonresponsive mutants have been extensively used as models for investigating the molecular mechanism of induction of CYP1A1 gene transcription by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. Incubation of cytosolic [3H]TCDD-aryl hydrocarbon (Ah) receptor from wild-type Hepa 1c1c7 cells for 16 h at 4 degrees C in 0.4 M KCl resulted in the formation of transformed liganded receptor which exhibited increased binding affinity on DNA-Sepharose columns. The elution properties of the peak with the highest DNA binding affinity were similar to the elution profiles of the nuclear receptor complex isolated from wild-type cells. TAOc1BPrcl (class I) nonresponsive mutant cells were characterized by relatively low levels of the cytosolic and nuclear Ah receptor complex. The BPrcl (class II) variant cell line contained levels of cytosolic receptor which were comparable to those observed in the wild-type cells; however, significantly reduced levels of nuclear receptor complex were observed in the class II variant cell line. Incubation of the nuclear or transformed liganded cytosolic Ah receptor from wild-type cells with a consensus 32P-labeled dioxin responsive element (DRE) in a gel shift assay gave a retarded band associated with the receptor-DRE complex. Incubation of the cytosolic receptor complex from the class I and II mutant cells for 16 h at 4 degrees C in 0.4 M KCl or for 2 h at 20 degrees C did not yield complexes with increasing binding affinities on DNA-Sepharose columns. Moreover, incubation of these complexes with 32P-labeled DRE did not give a retarded band in a gel shift assay. However, coincubation of the liganded class II mutant cytosol with cytosol from class I cells resulted in transformation of the liganded receptor and this was confirmed in both the DNA-Sepharose and gel retardation assays. These results suggest that the failure of class II mutant cells to respond to TCDD is due to a defect in the factors responsible for transformation of the cytosolic receptor complex.


Chemical structure of the lipid A component of lipopolysaccharides from Fusobacterium nucleatum.
The lipid A component of lipopolysaccharides from Fusobacterium nucleatum Fev 1 consists of beta-1',6-linked D-glucosamine disaccharides, which carry two phosphate groups: one in glycosidic and one in ester linkage. The amino groups of the glucosamine disaccharides are substituted by D-3-hydroxyhexadecanoic acid. The hydroxyl groups of the disaccharide backbone are acylated by tetradecanoic, hexadecanoic, and D-3-hydroxytetradecanoic acids. Part of the ester-bound D-3-hydroxytetradecanoic acid is 3-O-substituted by tetradecanoic acid. Whereas a similar pattern of fatty acids was detected in lipopolysaccharides from two other F. nucleatum strains, the amide-bound fatty acid in F. varium and F. mortiferum was D-3-hydroxytetradecanoic acid. The chemical relationships of lipid A from Fusobacteria and other gram-negative bacteria are discussed.


Taurine decreases fecal fatty acid and sterol excretion in cystic fibrosis. A randomized double-blind trial.
Patients with cystic fibrosis may still have a significant degree of steatorrhea despite adequate pancreatic enzyme supplementation. Taurine is a conditionally essential amino acid that possibly improves the micellar phase of fat digestion. Thirteen children with cystic fibrosis and a significant degree of steatorrhea (> 13 g/d) were enrolled in a randomized double-blind crossover study of taurine (30 mg/kg per day) in contrast to placebo for two successive 4-month periods. No difference was noted in height and weight velocity, lung function, vitamin A level, and essential fatty acid status. Twelve of the 13 patients showed a decrease in fecal fatty acid excretion (26.5 +/- 2.6 g/24 h vs 15.4 +/- 2.5 g/24 h), affecting mainly saturates and monounsaturates, and a decrease in total sterol excretion (1492.6 +/- 303 mg/24 h vs 1211.7 +/- 213.8 mg/24 h) while ingesting taurine. Taurine may be a useful adjunct in patients with cystic fibrosis and severe steatorrhea.


TRAIL promotes the survival and proliferation of primary human vascular endothelial cells by activating the Akt and ERK pathways.
TRAIL protein is expressed in the medial smooth cell layer of aorta and pulmonary artery, whereas endothelial cells express all TRAIL receptors (TRAIL-Rs). The role of TRAIL/TRAIL-Rs in vascular biology was investigated in primary human umbilical vein endothelial cells (HUVECs) and aortic endothelial cells, which showed comparable surface expression of death (TRAIL-R1 and -R2) and decoy (TRAIL-R3 and -R4) TRAIL-Rs. TRAIL activated the protein kinase Akt in HUVECs, as assessed by Western blot for phospho-Akt. Moreover, experiments performed with a pharmacological inhibitor of the phosphatidylinositol 3-kinase/Akt pathway (LY294002) or a dominant-negative Akt (K179M) demonstrated that TRAIL significantly protected HUVECs from apoptosis induced by trophic withdrawal via Akt and that inhibition of Akt sensitized HUVECs to TRAIL-induced caspase-dependent apoptosis. TRAIL also stimulated the ERK1/2 but not the p38 or the JNK pathways and induced a significant increase in endothelial cell proliferation in an ERK-dependent manner. Conversely, TRAIL did not activate NF-kappaB or affect the surface expression of the inflammatory markers E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1. The ability of TRAIL to promote the survival/proliferation of endothelial cells without inducing NF-kappaB activation and inflammatory markers suggests that the TRAIL/TRAIL-R system plays an important role in endothelial cell physiology.


Metal-Organic Framework-Based Nano-Activators Facilitating Microwave Combined Therapy via a Divide-and-Conquer Tactic for Triple-Negative Breast Cancer.
Aiming at the clinical problems of high recurrence and metastasis rate of triple-negative breast cancer, a divide-and-conquer tactic is developed. The designed nanoactivators enhance microwave thermo-dynamic-chemotherapy to efficiently kill primary tumors, simultaneously ameliorate the immunosuppressive microenvironment, activate the tumor infiltration of T lymphocytes, and enhance the accumulation and penetration of PD-1/PD-L1 immune agents, ultimately boosting the efficacy of immune checkpoint blocking therapy to achieve efficient inhibition of distal tumors and metastases. Metal-organic framework (MOF)-based MPPT nano-activator is synthesized by packaging chemotherapeutic drug Pyrotinib and immunosuppressant PD-1/PD-L1 inhibitor 2 into MnCa-MOF and then coupling target molecule triphenylphosphine, which significantly improved the accumulation and penetration of Pyrotinib and immunosuppressant in tumors. In addition to the combined treatment of microwave thermo-dynamic-chemotherapy under microwave irradiation, Mn2+ in the nano-activator comprehensively promotes the cGAS-STING pathway to activate innate immunity, microwave therapy, and hypoxia relief are combined to ameliorate the tumor immunosuppressive microenvironment. The released Pyrotinib down-regulates epidermal growth factor receptor and its downstream pathways PI3K/AKT/mTOR and MAPK/ERK signaling pathways to maximize the therapeutic effect of immune checkpoint blocking, which helps to enhance the antitumor efficacy and promote long-term memory immunity. This nano-activator offers a generally promising paradigm for existing clinical triple-negative breast cancer treatment through a divide-and-conquer strategy.


Carbon anhydrase IX specific immune responses in patients with metastatic renal cell carcinoma potentially cured by interleukin-2 based immunotherapy.
The majority of clear-cell renal cell carcinomas (ccRCC) show high and homogeneous expression levels of the tumor associated antigen (TAA) carbonic anhydrase IX (CAIX), and treatment with interleukin-2 (IL-2) based immunotherapy can lead to cure in patients with metastatic renal cell carcinoma (mRCC). However, the involvement of CAIX specific CD8+ T cells and/or NK cells in the tumor eradication is unknown. We investigated T cell and antibody reactivity against overlapping 15-mer CAIX-peptides as well as HLA haplotype frequency and NK cell cytotoxicity in 11 patients with no evidence of disease (NED) following treatment with IL-2 based immunotherapy, and thus potentially cured. Immune reactivity in these patients was compared with samples from patients with dramatic tumor response obtained immediately at the cessation of therapy, samples from patients that experienced progressive disease during treatment and samples from healthy controls. We observed more focused but only weak and not consistent CAIX specific T-cells in the late observation and early observation response groups compared with the healthy control group. An increased frequency of the class II alleles HLA-DRB4 01:01, HLA-DPB 01:01 and HLA-DPB 03:01 was noted in the NED patients. In contrast, NK cytotoxicity was low even in the late observation response group as compared with controls. In particular, a HLA-B*40:01 restricted CD8+ T cell response recognizing the CAIX- derived peptide SEEEGSLKL was identified. This may have interest in future cancer vaccines, but more studies are needed to elucidate the immunological mechanisms of action in potentially cured patients treated with an immunotherapeutic agent.


Micro-laser-induced breakdown spectroscopy technique: a powerful method for performing quantitative surface mapping on conductive and nonconductive samples.
Laser-induced breakdown spectroscopy (LIBS) has been applied mainly to bulk analysis of solids, liquids, and gases and less frequently for elemental microanalysis of solid surfaces. A micro-LIBS device devoted to analysis of the distribution of elements on surfaces is described. This device offers rapid access with a 3-microm spatial resolution to the microchemical structures of both conductive and nonconductive samples. Quantitative microchemical results of applications to ceramics are reported. By the use of a time-resolved acquisition spectrum, cerium in a uranium matrix was characterized with a cerium detection limit of 1.14%. Calibration curves obtained with manipulations during 1 year facilitated evaluations of reproducibility and repeatability. A 2% single-shot repeatability with a calibration reproducibility of approximately 7% is reported.


Complete Genome Sequence of Rhizobium japonicum Podophage Pasto.
Rhizobium japonicum is a Gram-negative bacterium of interest for research into nitrogen fixation in legumes. This article describes the isolation, sequencing, and annotation of R. japonicum podophage Pasto. While it shows no significant similarity to identified phages, genomic analysis indicates that Pasto may be temperate and is a novel T7-like podophage.


Contributions of the hippocampus and medial prefrontal cortex to energy and body weight regulation.
The effects of selective ibotenate lesions of the complete hippocampus (CHip), the hippocampal ventral pole (VP), or the medial prefrontal cortex (mPFC) in male rats were assessed on several measures related to energy regulation (i.e., body weight gain, food intake, body adiposity, metabolic activity, general behavioral activity, conditioned appetitive responding). The testing conditions were designed to minimize the nonspecific debilitating effects of these surgeries on intake and body weight. Rats with CHip and VP lesions exhibited significantly greater weight gain and food intake compared with controls. Furthermore, CHip-lesioned rats, but not rats with VP lesions, showed elevated metabolic activity, general activity in the dark phase of the light-dark cycle, and greater conditioned appetitive behavior, compared with control rats without these brain lesions. In contrast, rats with mPFC lesions were not different from controls on any of these measures. These results indicate that hippocampal damage interferes with energy and body weight regulation, perhaps by disrupting higher-order learning and memory processes that contribute to the control of appetitive and consummatory behavior.


A new electromechanical trainer for sensorimotor rehabilitation of paralysed fingers: a case series in chronic and acute stroke patients.
The functional outcome after stroke is improved by more intensive or sustained therapy. When the affected hand has no functional movement, therapy is mainly passive movements. A novel device for repeating controlled passive movements of paralysed fingers has been developed, which will allow therapists to concentrate on more complicated tasks. A powered cam shaft moves the four fingers in a physiological range of movement. After refining the training protocol in 2 chronic patients, 8 sub-acute stroke patients were randomised to receive additional therapy with the Finger Trainer for 20 min every work day for four weeks, or the same duration of bimanual group therapy, in addition to their usual rehabilitation. In the chronic patients, there was a sustained reduction in finger and wrist spasticity, but there was no improvement in active movements. In the subacute patients, mean distal Fugl-Meyer score (0-30) increased in the control group from 1.25 to 2.75 (ns) and 0.75 to 6.75 in the treatment group (p < .05). Median Modified Ashworth score increased 0/5 to 2/5 in the control group, but not in the treatment group, 0 to 0. Only one patient, in the treatment group, regained function of the affected hand. No side effects occurred. Treatment with the Finger Trainer was well tolerated in sub-acute & chronic stroke patients, whose abnormal muscle tone improved. In sub-acute stroke patients, the Finger Trainer group showed small improvements in active movement and avoided the increase in tone seen in the control group. This series was too small to demonstrate any effect on functional outcome however.


Nanostructure characterisation of flow-formed Cr-Mo-V steel using transmission Kikuchi diffraction technique.
Nowadays flow-forming has become a desired near net shape manufacturing method as it provides excellent mechanical properties with improved surface finish and significant manufacturing cost reduction. However, the material is subjected to excessive plastic deformation during flow-forming process, generating a very fine and complex microstructure. In addition, the intense dislocation density and residual stress that is generated in the component during processing makes the microstructure characterisation using conventional micro-analytical tools challenging. Thus, the microstructure/property relationship study in such a material is rather difficult. In the present study a flow-formed Cr-Mo-V steel nanostructure and crystallographic texture were characterised by means of Transmission Kikuchi Diffraction (TKD). Here, TKD is shown to be a powerful technique in revealing very fine martensite laths within an austenite matrix. Moreover, fine precipitates in the order of 20-70 nm on the martensite lath boundaries were clearly imaged and characterised. This greatly assisted in understanding the preferable site formation of the carbides in such a complex microstructure. The results showed that the actual TKD spatial resolution was in the range of 5-10 nm using 25 kV for flow-formed Cr-Mo-V steel.


How long should we continue crizotinib in ALK translocation-positive inflammatory myofibroblastic tumors? Long-term complete response with crizotinib and review of the literature.
Inflammatory myofibroblastic tumor is a rare disease which is typically seen in children and young adults. Approximately half of the inflammatory myofibroblastic tumors contain translocations that result in over-expression of anaplastic lymphoma kinase gene. Herein, we present two anaplastic lymphoma kinase-positive cases with long-term remission with crizotinib. We do not know how long these therapies need to be continued. We present two cases of inflammatory myofibroblastic tumor treated with anaplastic lymphoma kinase inhibitor therapies: an 8-year-old Turkish boy and a 21-year-old Caucasian man. Two cases, both with good tumor control under crizotinib, but one who progressed on drug holiday, responded again to the same drug, and had a very short period of response after restarting crizotinib. A molecular-targeted drug (anaplastic lymphoma kinase inhibitor) was found to be extremely effective as selective therapy for inflammatory myofibroblastic tumor with anaplastic lymphoma kinase translocation. Here, we want to emphasize the continuation of this treatment after achieving a good response until progression or a major side effect.


Rational design of modular polyketide synthases: morphing the aureothin pathway into a luteoreticulin assembly line.
The unusual nitro-substituted polyketides aureothin, neoaureothin (spectinabilin), and luteoreticulin, which are produced by diverse Streptomyces species, point to a joint evolution. Through rational genetic recombination and domain exchanges we have successfully reprogrammed the modular (type I) aur polyketide synthase (PKS) into a synthase that generates luteoreticulin. This is the first rational transformation of a modular PKS to produce a complex polyketide that was initially isolated from a different bacterium. A unique aspect of this synthetic biology approach is that we exclusively used genes from a single biosynthesis gene cluster to design the artificial pathway, an avenue that likely emulates natural evolutionary processes. Furthermore, an unexpected, context-dependent switch in the regiospecificity of a pyrone methyl transferase was observed. We also describe an unprecedented scenario where an AT domain iteratively loads an extender unit onto the cognate ACP and the downstream ACP. This aberrant function is a novel case of non-colinear behavior of PKS domains.


Postnatal pulmonary distension for the treatment of pulmonary hypoplasia: pilot study in the neonatal piglet model.
Accelerated lung growth has previously been demonstrated after fetal tracheal occlusion. The purpose of this study was to determine if short-term perfluorocarbon (PFC) distension could increase lung growth postnatally in neonatal piglets. Eleven piglets aged 5 to 8 days were divided into 3 groups: (a) controls (n = 4), (b) PFC x 6 hours (n = 3), and (c) PFC x 12 hours (n = 4). A right posterolateral thoracotomy was performed and a pressure-monitoring catheter was placed in the posterior segment of the right upper lobe. Perfluorocarbon was infused and a mean intrabronchial pressure of 12 mm Hg (range, 5-21 mm Hg) was maintained. The control piglets also had a thoracotomy with right upper lobe bronchus dissection without ligation or PFC distension. All piglets were injected with [3H]-thymidine 3 hours before killing. Both right and left posterior segments of each upper lobe were analyzed for their respective amount of total DNA by fluorometry. Rates of DNA synthesis for each segment were determined by precipitating incorporated [3H]-thymidine with 5% trichloroacetic acid and reporting this value by the total amount of DNA. The differential lung DNA synthesis rate was calculated as (right posterior segment/left posterior segment) x 100. Statistical analysis consisted of 1-way analysis of variance and Student's t tests with significance at P < or = .05. Heart rate, mean arterial pressure, temperature, oxygen saturation, pH, PCO2 , and PO2 were similar in all 3 groups. Lung DNA synthesis was nearly doubled in the PFC x 6 hours group compared with controls (302% vs 165%, P = .05). Animals in the PFC x 12 hours group experienced a 261% increase (P = NS). Short-term PFC distension in neonatal piglets resulted in increased DNA synthesis within 6 hours presumably because of stretch-induced mechanisms.


A longitudinal examination of the interpersonal theory of suicide and effects of school-based suicide prevention interventions in a multinational study of adolescents.
Predictions of two different models for suicide attempts (SA) over 12 months, as differentially impacted by specific school-based suicide prevention interventions, were compared. These were as follows: (a) interpersonal theory (IPTS) and (b) a two-pathway model, one path associated with externalizing symptoms and continuum of self-harm behaviors, and the other with internalizing symptoms. Self-report questionnaires were completed by 11,110 high school students from ten EU countries enrolled in the Saving and Empowering Young Lives in Europe (SEYLE) study. Baseline measures included perceived burdensomeness, thwarted belongingness from parents and peers, health risk behaviors, self-injury, suicide ideation (SI), and attempts (SA). SI and SA were reassessed at 12-month follow-up. Each model's predictions of SI and SA groups over time (i.e., repeated SA, remitted SA, SA onset, and no SA) were estimated in the no intervention/control group. The superior model was estimated across intervention groups. Interpersonal theory showed better fit compared to the two-pathway model. In partial agreement with IPTS predictions, parental low belongingness but not peer belongingness or burdensomeness predicted greater likelihood of SI. The likelihood of repeated SA versus no SA was higher among adolescents who reported SI, self-injury, risk behaviors, and particularly both SI and self-injury. All three interventions attenuated the combined effect of SI and self-injury. Youth Aware of Mental Health Program (YAM) additionally decreased the effect of risk behaviors on the likelihood of repeated SA. Interpersonal theory assumptions were partially supported. Perceived interpersonal difficulties with parents were primarily related with SI, and risk behaviors and self-injury were important predictors of SA. Suicide prevention interventions may be effective by mitigating the hazardous effect of varying self-harm behaviors and may be further advanced by increasing parental involvement.


The genome of the obligate endobacterium of an AM fungus reveals an interphylum network of nutritional interactions.
As obligate symbionts of most land plants, arbuscular mycorrhizal fungi (AMF) have a crucial role in ecosystems, but to date, in the absence of genomic data, their adaptive biology remains elusive. In addition, endobacteria are found in their cytoplasm, the role of which is unknown. In order to investigate the function of the Gram-negative Candidatus Glomeribacter gigasporarum, an endobacterium of the AMF Gigaspora margarita, we sequenced its genome, leading to an ∼1.72-Mb assembly. Phylogenetic analyses placed Ca. G. gigasporarum in the Burkholderiaceae whereas metabolic network analyses clustered it with insect endobacteria. This positioning of Ca. G. gigasporarum among different bacterial classes reveals that it has undergone convergent evolution to adapt itself to intracellular lifestyle. The genome annotation of this mycorrhizal-fungal endobacterium has revealed an unexpected genetic mosaic where typical determinants of symbiotic, pathogenic and free-living bacteria are integrated in a reduced genome. Ca. G. gigasporarum is an aerobic microbe that depends on its host for carbon, phosphorus and nitrogen supply; it also expresses type II and type III secretion systems and synthesizes vitamin B12, antibiotics- and toxin-resistance molecules, which may contribute to the fungal host's ecological fitness. Ca. G. gigasporarum has an extreme dependence on its host for nutrients and energy, whereas the fungal host is itself an obligate biotroph that relies on a photosynthetic plant. Our work represents the first step towards unraveling a complex network of interphylum interactions, which is expected to have a previously unrecognized ecological impact.


Functional Incidental Training: applicability and feasibility in the Veterans Affairs nursing home patient population.
To examine the applicability and feasibility of an intervention directed at improving continence, endurance, and strength (Functional Incidental Training [FIT]), for older patients in Veterans Administration (VA) nursing homes. Data were collected during a randomized, controlled, crossover trial. Four VA nursing homes. All 528 patients in the nursing homes were screened, 178 were eligible, and 107 were randomized into the trial. A total of 64 participants completed the intervention phase of the trial. Trained research staff provided the FIT intervention, which included prompted voiding combined with individualized, functionally oriented endurance and strength training exercises offered four times per day, 5 days per week, for 8 weeks. Descriptive data were collected relevant to the translation of the FIT intervention into everyday practice, including number of patients eligible and reasons for ineligibility, attrition rates and reasons for attrition, participant adherence to and satisfaction with FIT, and the costs of FIT relative to usual care. One third of the 528 patients met the eligibility criteria. The major reasons for ineligibility were being continent, age under 60, and a short anticipated length of stay. Of the 146 patients enrolled in the trial, 85 (58%) dropped out during the 9- to 10-month project. Deterioration in health status, death, and discharge accounted for two thirds of the attrition. Adherence to FIT was in general high but variable. Participants completed prompted voiding plus at least one exercise in 75% of the FIT rounds offered. Of the 60 participants who completed the protocol and who could answer simple questions, 75% indicated they enjoyed FIT, but 62% indicated that the exercise was too frequent, and 28% indicated they were offered opportunities to toilet too often. Based on timed observations, the costs of FIT are about four times as high as usual continence care. FIT is applicable to a substantial number of patients in VA nursing homes. The FIT protocol tested in this trial can be further refined and individualized based on patient preferences and adherence to various components of FIT in order to make it more feasible, efficient, and cost-effective in practice. The costs of maintaining an intervention such as FIT in all VA nursing home patients who may benefit, however, are high and must be justified largely by potential positive effects on function and quality of life, as opposed to cost savings resulting from the intervention.


Whole blood viscosity parameters and cerebral blood flow.
This report describes the statistical relationship of several whole blood viscosity parameters and cerebral blood flow (CBF) in 53 consecutive patients and normal controls. Significant correlations were present between CBF and serum fibrinogen (P = .05), hematocrit (P less than .05), and a relationship involving both fibrinogen and hematocrit (P less than .01). We conclude that heightened whole blood viscosity does correlate with decreased cerebral blood flow in the ranges measured in our patients, that both fibrinogen and hematocrit must be taken into consideration in viscosity determinations, and that changes in viscosity may have an important effect on CBF in regions of low flow.


Management of end-stage heart failure patients with or without ventricular assist device: an observational comparison of clinical and economic outcomes.
Heart transplantation (HT) and ventricular assist devices (VAD) for the management of end-stage heart failure have not been directly compared. We compare the outcomes and use of resources with these 2 strategies in 2 European countries with different allocation systems. We studied 83 patients managed by VAD as the first option in Bad Oeynhausen, Germany (Group I) and 141 managed with either HT or medical therapy, as the first option, in Paris, France (Group II). The primary end-point was 2-year survival. Kaplan-Meier analyses were performed after the application of propensity score weights to mitigate the effects of non-random group assignment. The secondary end-points were resource utilization and costs. Subgroup analyses were performed for patients undergoing HT and patients treated with inotropes at the enrolment time. The Group I patients were more severely ill and haemodynamically compromised, and 28% subsequently underwent HT vs 55% primary HT in Group II, P < 0.001. The adjusted probability of survival was 44% in Group I vs 70% in Group II, P <0.0001. The mean cumulated 2-year costs were €281 361 ± 156 223 in Group I and €47 638 ± 35 061 in Group II, P < 0.0001. Among patients who underwent HT, the adjusted probability of survival in Group I (n = 23) versus Group II (n = 78) was 76% versus 68%, respectively (0.09), though it differed in the inotrope-treated subgroups (77% in Group I vs 67% in Group II, P = 0.04). HT should remain the first option for end-stage heart failure patients, associated with improved outcomes and better cost-effectiveness profile. VAD devices represent an option when transplant is not possible or when patient presentation is not optimal.


Customization of UWB 3D-RTLS Based on the New Uncertainty Model of the AoA Ranging Technique.
The increased potential and effectiveness of Real-time Locating Systems (RTLSs) substantially influence their application spectrum. They are widely used, inter alia, in the industrial sector, healthcare, home care, and in logistic and security applications. The research aims to develop an analytical method to customize UWB-based RTLS, in order to improve their localization performance in terms of accuracy and precision. The analytical uncertainty model of Angle of Arrival (AoA) localization in a 3D indoor space, which is the foundation of the customization concept, is established in a working environment. Additionally, a suitable angular-based 3D localization algorithm is introduced. The paper investigates the following issues: the influence of the proposed correction vector on the localization accuracy; the impact of the system's configuration and LS's relative deployment on the localization precision distribution map. The advantages of the method are verified by comparing them with a reference commercial RTLS localization engine. The results of simulations and physical experiments prove the value of the proposed customization method. The research confirms that the analytical uncertainty model is the valid representation of RTLS' localization uncertainty in terms of accuracy and precision and can be useful for its performance improvement. The research shows, that the Angle of Arrival localization in a 3D indoor space applying the simple angular-based localization algorithm and correction vector improves of localization accuracy and precision in a way that the system challenges the reference hardware advanced localization engine. Moreover, the research guides the deployment of location sensors to enhance the localization precision.


Exploring the Biomedical Potential of PLA/Dysprosium Phosphate Composites via Extrusion-Based 3D Printing: Design, Morphological, Mechanical, and Multimodal Imaging and Finite Element Modeling.
The present investigation demonstrates the feasibility of dysprosium phosphate (DyPO4) as an efficient additive in polylactide (PLA) to develop 3D printed scaffolds through the material extrusion (MEX) principle for application in bone tissue engineering. Initially, uniform sized particles of DyPO4 with tetragonal crystal setting are obtained and subsequently blended with different concentrations of PLA to extrude in the form of filaments. A maximum of 20 wt % DyPO4 in PLA matrix has been successfully drawn to yield a defect free filament. The resultant filaments were 3D printed through material extrusion methodology. The structural and morphological analysis confirmed the successful reinforcement of DyPO4 throughout the PLA matrix in all of the 3D printed components. All of the PLA/DyPO4 composites exhibited magnetic resonance imaging and computed tomography contrasting properties, which were dependent on the dysprosium content in the PLA matrix. The detailed mechanical evaluation of the 3D printed PLA/DyPO4 composites ensured good strength accomplished by the reinforcement of 5 wt % DyPO4 in PLA matrix, beyond which a gradual decline in the strength is noticed. Representative volume elements models were developed to realize the intrinsic property of the PLA/DyPO4 composite, and finite element analysis under both static and dynamic loading conditions has been performed to account for the reliability of experimental results.


Preneoplastic changes persist after IGF-IR downregulation and tumor regression.
Despite our incomplete understanding of the function of the type I insulin-like growth factor receptor (IGF-IR) in tumorigenesis, IGF-IR targeting agents have entered clinical trials for the treatment of human cancers. Previously, we have shown that downregulation of IGF-IR transgene in mammary tumors in MTB-IGFIR transengic mice results in tumor regression in a majority of the mice and most of these mice do not develop recurrent mammary tumors. In this study, we examined mammary tissue of mice that did not develop recurrent tumors. Areas of tumor regression were visible macroscopically and microscopically these lesions contained cell debris, individual cells, lipofuscin and doxycycline crystals. Three of the 12 mice also presented with considerable lobuloalveolar development. The re-expression of the IGF-IR transgene in mammary tissue with stably regressed tumors resulted in the rapid re-emergence of mammary tumors, some of which seemed to originate from the regressed mammary lesions. Thus, despite stable tumor regression after IGF-IR downregulation, mammary tissue contained preneoplastic lesions and tumors rapidly re-appear upon re-overexpression of IGF-IR transgene. Therefore, IGF-IR-targeting agents may be effective at regressing mammary tumors expressing IGF-IR, but these agents will not completely eradicate all tumor cells or restore the mammary stromal environment.


Organ-resident, nonlymphoid cells suppress proliferation of autoimmune T-helper lymphocytes.
Local presentation of autoantigen by organ-resident cells inappropriately expressing Ia determinants has been implicated in organ-specific autoimmunity. Experimental autoimmune uveoretinitis, induced in rats by immunization with retinal soluble antigen, is used as a model of organ-specific autoimmunity. In an in vitro system derived from this model, uveitogenic rat T-helper lymphocytes specific to the retinal soluble antigen, or control T-helper lymphocytes reactive to the purified protein derivative of tuberculin, were cocultured with Ia-expressing syngeneic retinal glial cells (Müller cells) in the presence of specific antigen. Antigen presentation was not apparent under ordinary culture conditions, and the Müller cells profoundly suppressed the proliferative response of primed T-helper lymphocytes to antigen presented on conventional antigen-presenting cells, as well as their subsequent interleukin-2 (IL-2)-dependent expansion. Suppression of proliferation was accompanied by inhibition of IL-2 production in response to antigen, as well as by reduction in high-affinity IL-2 receptor expression, and proceeded via a contact-dependent mechanism. These results suggest a role for locally acting suppression mechanisms in immune regulation and maintenance of tissue homeostasis.


[The characteristics of the work process of dental technicians and their body functional status in the dynamics of work activities].
The article presents in the ophthalmoergonomic aspect data from studies on the character of labour activity, condition of visual organs, vision conditions, and functions of the visual analyzer in dental mechanics. It has been ascertained that during the labour process, it is the functions of accommodation and contrast sensitivity of the eye that bear the highest strain (P < 0.05), and by the vision conditions delineation, the dental mechanic's work is on the list of those jobs requiring high visual efficiency (III B category of visual jobs according to the adopted and enforced standards classification), which facts necessitate ensuring high levels of illumination. Recommendatory measures are presented designed to prevent in dental mechanics general and visual fatigue (eyestrain and to maintain their capacity for work.


Monoclonal immunofluorescence compared with silver stain for investigating Pneumocystis carinii pneumonia.
Two hundred and eighty two specimens from 220 patients positive for HIV with respiratory tract symptoms, or febrile illness, or both, were examined for the presence of Pneumocystis carinii. Specimens were either induced sputum samples or bronchoalveolar lavage fluids. To establish the optimal method for laboratory diagnosis a comparison was made of detection of the organism by use of monoclonal antibody and immunofluorescence with conventional silver staining methods. Three commercially available reagents for immunofluorescence were also compared. Immunofluorescence was significantly more sensitive than the silver stain and the best results for immunofluorescence were obtained using. Northumbria Biologicals Ltd reagents.


Ex vivo lung perfusion with perfusate purification for human donor lungs following prolonged cold storage.
Ex vivo lung perfusion (EVLP) has developed as the most effective technique for estimating marginal donor lungs. This study attempted to extend the EVLP running time to 12 hours with a dialyzer rather than periodically replacing the perfusate. Human donor lungs rejected by the clinical lung transplantation (LTx) team were obtained. After cold storage lasting 18-24 hours, lungs were randomly assigned to 2 groups: a control group and a perfusate purification (PP) group. The control group underwent EVLP in the traditional way, while a dialyzer was added into the circuit as a bypass in the PP group. The effects on lung function, microenvironment, inflammatory response, and cell death were evaluated. A total of 8 rejected donor lungs were obtained and each group was assigned to 4 cases of EVLP. Three cases were prematurely terminated because of serious lung edema and decreased lung function. There were no significant differences in airway pressure, pulmonary artery pressure, and oxygen concentration between the two groups in the first 8 hours. The pH in the control group was significantly lower than that in the PP group, and the levels of potassium and lactate were significantly higher than those in the PP group. Inflammatory markers increased in both groups, while IL-6 and IL-10 were higher in the PP group in the first 6 hours. Hematoxylin and eosin (HE) staining revealed lung injuries in both groups, but no significant difference was noted in the HE-stained slides. There were significantly more TUNEL-positive cells in the control group (69.5%±4.0%) than in the PP group (47.5%±3.9%) (P=0.000). Using the modified method of EVLP reduces the high cost caused by exchanging perfusion fluid per hour and could prolong the normothermic preservation time of donor lungs.


Culturing and Cryobanking Human Neural Stem Cells.
The discovery and study of human neural stem cells has advanced our understanding of human neurogenesis, and the development of novel therapeutics based on neural cell replacement. Here, we describe methods to culture and cryopreserve human neural stem cells (hNSCs) for expansion and banking. Importantly, the protocols ensure that the multipotency of hNSCs is preserved to enable differentiation to neurons and supporting neuroglia.


Use of primary cultures of adult rat hepatocytes on collagen gel-nylon mesh to evaluate carcinogen-induced unscheduled DNA synthesis.
The procarcinogen, 2-acetylaminofluorene, the direct-acting carcinogen, methyl methanesulfonate, and two other hepatocarcinogens, thioacetamide and urethan, were tested for their ability to elicit unscheduled DNA synthesis in adult rat hepatocytes maintained in primary culture on collagen gel-nylon mesh. The carcinogens, dissolved in dimethyl sulfoxide were added to 6-hr or to 28-hr cultures along with [methyl-3H]thymidine (1 muCi/ml medium) in the presence of 10 mM hydroxyurea. Twelve hr later, the hepatocytes were harvested from the cultures with collagenase, and their DNA was purified on CsCl isopyknic gradients. Unscheduled DNA synthesis was measured as the increase in [methyl-3H]thymidine radioactivity incorporated per microgram DNA of the carcinogen-treated cultures as compared with that of control cultures. Both 2-acetylaminofluorene and methyl methanesulfonate demonstrated a concentration-dependent stimulation of unscheduled DNA synthesis in the 6-hr hepatocyte cultures. However, the response of the 28-hr cultures to these two carcinogens was absent unless the hepatocytes were preincubated for 22 hr in culture medium supplemented with 10(-5) M dexamethasone and 10(-6) M glucagon or in a more complete hormone-supplemented medium. Thioacetamide and urethan, on the other hand, failed to elicit a concentration-dependent unscheduled DNA synthesis under these conditions. The results obtained with this culture system are similar to those of other short-term tests for chemical carcinogenicity and support the potential use of the collagen gel-nylon mesh-hepatocyte primary culture as an in vitro screen for chemical carcinogens. Furthermore, this study suggests the importance of specific hormones in maintaining the capability for repair of DNA damage produced by carcinogenic and mutagenic chemicals in cultured hepatocytes.


Automated estimated GFR reporting: A new tool to promote safer prescribing in patients with chronic kidney disease?
A number of drugs rely on the kidney for excretion and thus require their dose to be modified in any patients where there is renal impairment. Others are nephrotoxic and should be avoided completely in patients with renal disease. Traditionally clinicians have had to rely on serum creatinine to assess renal function but this may not accurately reflect the kidney function of an individual since its level also depends on muscle mass. In particular elderly females may have significant impairment of renal function despite a normal or near normal serum creatinine. The advent of automated reporting of estimated glomerular filtration rate (eGFR) provides the clinician with simple, easily understood and readily available measurement which more accurately reflects a patient's renal function. In particular eGFR allows the clinician to readily identify and stratify patients with chronic kidney disease (CKD) and can allow a more rational and safer approach to prescribing in this group of high risk patients. This commentary suggests that national prescribing formularies should use eGFR to provide consistent advice about the appropriate dose adjustment and avoidance of potentially toxic drugs at various stages of CKD. Such an approach may prove invaluable in improving prescribing in CKD and avoiding drug toxicity in this group of patients.


ADAR activation by inducing a syn conformation at guanosine adjacent to an editing site.
ADARs (adenosine deaminases acting on RNA) can be directed to sites in the transcriptome by complementary guide strands allowing for the correction of disease-causing mutations at the RNA level. However, ADARs show bias against editing adenosines with a guanosine 5' nearest neighbor (5'-GA sites), limiting the scope of this approach. Earlier studies suggested this effect arises from a clash in the RNA minor groove involving the 2-amino group of the guanosine adjacent to an editing site. Here we show that nucleosides capable of pairing with guanosine in a syn conformation enhance editing for 5'-GA sites. We describe the crystal structure of a fragment of human ADAR2 bound to RNA bearing a G:G pair adjacent to an editing site. The two guanosines form a Gsyn:Ganti pair solving the steric problem by flipping the 2-amino group of the guanosine adjacent to the editing site into the major groove. Also, duplexes with 2'-deoxyadenosine and 3-deaza-2'-deoxyadenosine displayed increased editing efficiency, suggesting the formation of a Gsyn:AH+anti pair. This was supported by X-ray crystallography of an ADAR complex with RNA bearing a G:3-deaza dA pair. This study shows how non-Watson-Crick pairing in duplex RNA can facilitate ADAR editing enabling the design of next generation guide strands for therapeutic RNA editing.


[Factors associated to glycaemic control in patients with type 2 diabetes].
Evaluate the factors influencing blood glucose control of type 2 diabetic patients attended at a tertiary health care center. A retrospective study by review of medical records of patients who attended the Diabetes Clinic at the University of Caxias do Sul was carried out. Patients were evaluated for glycaemic and metabolic control and divided according to glycated hemoglobin at the beginning and end of the period in question, in compensated and not compensated. The factors associated with glycaemic control were analyzed. Seventy three patients were included in the analysis. On the average, improvement of parameters related to glycaemic, pressure and lipid control during the average of 20 months of treatment was observed. In the beginning of the follow-up period, 25% of patients had glycated hemoglobin < 7%, 22.7% had LDL cholesterol < 100mg/dL, 8.7% had systolic blood pressure < 130 mmHg and diastolic < 80 mmHg and no patients had all parameters meeting the recommended goals. At the last visit the percentages were, 42.3%, 37.5%, 30.2% and 9.6% respectively. In the multivariate analysis, younger age, higher levels of LDL cholesterol and insulin use were associated with poor blood glucose control. A small part of the sample simultaneously achieved the goals for blood glucose, lipids and blood pressure control. Some factors, such as age and gender can be determinants to achieve control of diabetes. Greater efforts are still needed to achieve the goals for treatment in type 2 diabetic patients.